PRIMJENA MULTIPLEKS LANČANE REAKCIJE POLIMERAZOM ZA DOKAZIVANJE TCDA I TCDB GENA ZA TOKSINE BAKTERIJE CLOSTRIDIOIDES DIFFICILE U FECESU PASA

master's thesis

• Title (english): Performance of multiplex polymerase chain reaction for detection of tcdA and tcdB genes of Clostridioides difficile toxins in feces of dogs
• Author: Magda Kamber
• Mentor: Suzana Hađina (mentor)
• Committee member: Josipa Habuš (predsjednik povjerenstva)
• Committee member: Zrinka Štritof (član povjerenstva)
• Committee member: Suzana Hađina (član povjerenstva)
• Committee member: Vilim Starešina (član povjerenstva)
• Granter: University of Zagreb; Faculty of Veterinary Medicine; Zagreb
• Defense date and country: 2022-08, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Veterinary Medicine; Veterinary Public Health and Food Safety

Abstract

Clostridioides difficile je gram-pozitivna, anaerobna, štapićasta, sporulirajuća bakterija
koja proizvodi glavna dva toksina: toksin A i toksin B čija je sinteza kodirana tcdA odnosno
tcdB genom. Smatra se da toksogeni sojevi dovode do nastanka različitih kliničkih znakova
bolesti probavnog sustava, od blagih samoograničavajućih proljeva pa sve do teških oblika
hemoragičnih gastroenteritisa. Cilj ovog istraživanja bio je utvrditi prisutnost gena tcdA i tcdB
za toksine bakterije Clostridioides difficile u fecesu zdravih pasa i pasa s kliničkim znakovima
bolesti probavnog sustava koristeći multipleks lančanu reakciju polimerazom te njihovu ulogu
u nastanku probavnih poremećaja u pasa. U ovom istraživanju analizirano je 116 uzoraka fecesa
pasa, odnosno 49 uzoraka fecesa zdravih pasa te 67 uzoraka fecesa pasa s kliničkim znakovima
bolesti probavnog sustava. U 5,2% uzoraka utvrđena je prisutnost tpi gena bakterije C. difficile,
od kojih je u 2,6% utvrđen toksogeni soj (A+B-), a u njih 2,6% netoksogeni soj (A-B-). U
zdravih životinja utvrđena je prisutnost tpi gena, odnosno netoksogeni soj bakterije C. difficile
(A-B-). U tri psa s kliničkim znakovima bolesti probavnog sustava utvrđena je prisutnost tcdA
gena za toksin A, odnosno toksogeni soj (A+B-), što ukazuje na moguću povezanost toksogenih
sojeva s nastankom kliničkih znakova bolesti probavnog sustava, dok je u jednog psa iz te
skupine utvrđen netoksogeni soj (A-B-). Rezultati ovog istraživanja ukazuju na potrebu
daljnjeg rasvjetljavanja prisutnosti i uloge bakterije C. difficile u nastanku bolesti probavnog
sustava u pasa.

Abstract (english)

Clostridioides difficile is a gram-positive, anaerobic, rod-shaped, sporulating bacterium
that produces two main types of toxins: toxin A and toxin B whose production is encoded by
the tcdA and tcdB genes. Toxigenic strains are thought to lead to the occurrence of various
gastrointestinal symptoms, ranging from mild self-limiting diarrhea to severe forms of
hemorrhagic gastroenteritis. The aim of this study was to determine the presence of the tcdA
and tcdB genes for C. difficile toxins in the feces of healthy dogs and dogs with symptoms of
gastrointestinal disease using multiplex polymerase chain reaction, and to determine their role
in the development of digestive disorders in dogs. In this study, 116 dog fecal samples were
analyzed, i.e., 49 fecal samples from healthy dogs and 67 fecal samples from dogs with
gastrointestinal symptoms. The tpi gene was detected in 5.2% of the fecal samples, with the
toxigenic strain (A+B-) detected in 2.6%, and the non-toxigenic strain (A-B-) detected in 2.6%
of the samples. In healthy animals, only the tpi gene was detected, i.e., a non-toxigenic strain
of C. difficile (A-B-). In three dogs with symptoms of gastrointestinal disease the tcdA gene of
toxin A was detected, i.e., a toxigenic strain (A+B+), indicating a possible association between
toxigenic strains and the occurrence of gastrointestinal symptoms, while the sample of one dog
from this group was identified as a non-toxigenic strain (A-B-). The results of this study indicate
that further research is needed on the presence and role of C. difficile in the occurrence of
gastrointestinal disease in dogs.

• Keywords: Clostridioides difficile; tcdA i tcdB geni za toksine A i B; multipleks lančana reakcija polimerazom; pas; bolesti probavnog sustava
• Keywords (english): Clostridioides difficile; tcdA and tcdB genes for toxins A and B; mutiplex polymerase chain reaction; dog; gastrointestinal disease
• Language: croatian
• URN:NBN: urn:nbn:hr:178:636900
• Study programme: Title: Veterinary Medicine; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: doktor/doktorica veterinarske medicine (doktor/doktorica veterinarske medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2022-09-12 11:18:03